Clin-Chem Lab AA Part 3

studied byStudied by 0 people
0.0(0)
learn
LearnA personalized and smart learning plan
exam
Practice TestTake a test on your terms and definitions
spaced repetition
Spaced RepetitionScientifically backed study method
heart puzzle
Matching GameHow quick can you match all your cards?
flashcards
FlashcardsStudy terms and definitions

1 / 29

encourage image

There's no tags or description

Looks like no one added any tags here yet for you.

30 Terms

1

Principle:

Measures total protein based on nitrogen content

KJELDAHL

New cards
2

What is the Reaction of KJELDAHL method.

• (Proteins + sulfuric acid)

• Nitrogen → Ammonium Sulfate → Distillation into Ammonia → Titration

New cards
3

What are the Disadvantages of KJELDAHL?

• Time-consuming, labor- intensive.

• Cannot differentiate protein nitrogen from nonprotein nitrogen.

New cards
4

Three (3) steps of KJELDAHL method?

(1) Digestion

(2) Neutralization and Distillation

(3) Titration

<p>(1) Digestion</p><p>(2) Neutralization and Distillation</p><p>(3) Titration</p>
New cards
5

Formula of (1) Digestion

Proteins + H2SO4→(NH4)2SO4

New cards
6

In this step, Serum proteins are precipitated with an organic acid (Trichloroacetic acid or tungstic acid)

(1) Digestion

New cards
7

In (1) Digestion, the protein pellet is digested in:

Sulfuric acid (H2SO4) with heat (340°C to 360°C)

New cards
8

This is the Catalyst in (1) Digestion.

Cupric sulfate (to speed up the reaction)

New cards
9

This is used to increase the boiling point in (1) Digestion.

Potassium sulfate (improves the efficiency of digestion).

New cards
10

Formula of (2) Neutralization and Distillation

(NH4)2SO4+2NaOH→2NH3+Na2SO4+2H2O

New cards
11

In this step, the digested solution is neutralized with a strong base

(2) Neutralization and Distillation

New cards
12

What is the strong base used in (2) Neutralization and Distillation?

NaOH (sodium hydroxide).

New cards
13

In this step, ammonia (NH₃) gas is released, which is distilled into a receiving solution containing boric acid (H₃BO₃), forming ammonium borate.

(2) Neutralization and Distillation

New cards
14

In this step, the ammonium borate (NH4H2BO3) formed is then titrated with a standard solution of HCl to determine the amount of nitrogen in the original protein solution.

(3) Titration

New cards
15

Final calculation in KJELDAHL method:

Total Protein (g/dL)=Nitrogen (g/dL)×6.25

<p>Total Protein (g/dL)=Nitrogen (g/dL)×6.25</p>
New cards
16

Why do we multiply the Nitrogen content by 6.25, in determining the Total Protein in KJELDAHL method?

The factor 6.25 is derived from the assumption that proteins are 16% nitrogen: 100% ÷ 16% = 6.25

New cards
17

Principle:

Cupric ions (Cu²⁺) react with peptide bonds in an alkaline environment to form a violet-colored complex

BIURET method.

New cards
18

• Most widely used method

• Recommended by IFCC for determination of TP

BIURET method.

New cards
19

Reaction that takes place in BIURET method:

Cu²⁺ + Peptide bonds → Violet color (measured at 540 nm)

New cards
20

Disadvantages of BIURET method:

Hemoglobin and lipemia interference

New cards
21

What are the components of BIURET REAGENT?

Potassium hydroxide

Sodium potassium tartrate

Potassium iodide

New cards
22

Under Biuret Reagent this:

Provides an alkaline medium so that the reaction can take place

Potassium hydroxide

New cards
23

Under Biuret Reagent this:

Contains Complex cupric ions

Sodium potassium tartrate

New cards
24

Under Biuret Reagent this:

Acts as an antioxidant.

Potassium iodide

New cards
25

Biuret Reagent’s absorbance is measured at?

540 nm

New cards
26

Principle:

• Proteins bind to certain dyes, causing color change proportional to protein concentration

• Used to stain protein bands after electrophoresis.

DYE-BINDING

New cards
27

Common dyes used in DYE-BINDING (Total Proein):

• Bromophenol blue

• Ponceau S

• Amido black 10B

• Lissamine green

• Coomassie brilliant blue

New cards
28

In Dye-Binding (Total Proein):

Relies on the binding to protein

Coomassie brilliant blue 250

New cards
29

Coomassie brilliant blue 250 has a shift absorbance maximum of the dye from?

465 to 595 nm

New cards
30

In Coomassie brilliant blue 250, absorbance at 595 nm = ?

protein concentration

New cards

Explore top notes

note Note
studied byStudied by 98 people
693 days ago
5.0(4)
note Note
studied byStudied by 241 people
678 days ago
5.0(2)
note Note
studied byStudied by 13 people
833 days ago
5.0(1)
note Note
studied byStudied by 52 people
787 days ago
5.0(1)
note Note
studied byStudied by 33 people
425 days ago
5.0(1)
note Note
studied byStudied by 9 people
169 days ago
5.0(2)
note Note
studied byStudied by 41 people
709 days ago
5.0(1)
note Note
studied byStudied by 100 people
760 days ago
5.0(2)

Explore top flashcards

flashcards Flashcard (112)
studied byStudied by 55 people
302 days ago
5.0(2)
flashcards Flashcard (198)
studied byStudied by 1 person
182 days ago
5.0(1)
flashcards Flashcard (24)
studied byStudied by 34 people
759 days ago
5.0(1)
flashcards Flashcard (47)
studied byStudied by 1 person
39 days ago
5.0(1)
flashcards Flashcard (137)
studied byStudied by 301 people
567 days ago
4.5(4)
flashcards Flashcard (308)
studied byStudied by 7 people
832 days ago
5.0(1)
flashcards Flashcard (35)
studied byStudied by 2 people
303 days ago
5.0(1)
flashcards Flashcard (334)
studied byStudied by 88 people
23 days ago
5.0(2)
robot